Bioengineering Laboratory, Department of Chemical and Biological Engineering, University at Buffalo, State University of New York, Amherst, NY 14260, United States.
J Control Release. 2010 Jun 1;144(2):213-20. doi: 10.1016/j.jconrel.2010.02.009. Epub 2010 Feb 11.
We employed fibrin hydrogel as a bioactive matrix for lentivirus mediated gene transfer. Fibrin-mediated gene transfer was highly efficient and exhibited strong dependence on fibrinogen concentration. Efficient gene transfer was achieved with fibrinogen concentration between 3.75 and 7.5mg/ml. Lower fibrinogen concentrations resulted in diffusion of virus out of the gel while higher concentrations led to ineffective fibrin degradation by target cells. Addition of fibrinolytic inhibitors decreased gene transfer in a dose-dependent manner suggesting that fibrin degradation by target cells may be necessary for successful gene delivery. Under these conditions transduction may be limited only to cells interacting with the matrix thereby providing a method for spatially-localized gene delivery. Indeed, when lentivirus-containing fibrin microgels were spotted in an array format gene transfer was confined to virus-containing fibrin spots with minimal cross-contamination between neighboring sites. Collectively, our data suggest that fibrin may provide an effective matrix for spatially-localized gene delivery with potential applications in high-throughput lentiviral microarrays and in regenerative medicine.
我们采用纤维蛋白水凝胶作为介导慢病毒基因转移的生物活性基质。纤维蛋白介导的基因转移非常高效，并表现出对纤维蛋白原浓度的强烈依赖性。纤维蛋白原浓度在 3.75 至 7.5mg/ml 之间时，可实现高效的基因转移。较低的纤维蛋白原浓度会导致病毒从凝胶中扩散，而较高的浓度则会导致靶细胞无法有效降解纤维蛋白。纤维蛋白溶解抑制剂的添加以剂量依赖的方式降低了基因转移，这表明靶细胞对纤维蛋白的降解可能是成功基因传递所必需的。在这些条件下，转导可能仅限于与基质相互作用的细胞，从而为空间定位基因传递提供了一种方法。事实上，当含有慢病毒的纤维蛋白微凝胶以阵列形式点样时，基因转移仅限于含有病毒的纤维蛋白点，相邻位点之间的交叉污染最小。总的来说，我们的数据表明纤维蛋白可能为空间定位基因传递提供一种有效的基质，在高通量慢病毒微阵列和再生医学中有潜在的应用。
